Single cell transcriptomic analysis of renal allograft rejection reveals novel insights into intragraft TCR clonality.
Tiffany ShiAshley R BurgJ Timothy CaldwellKrishna RoskinCyd M Castro-RojasP Chukwunalu ChukwumaGeorge I GraySara G FooteJesus AlonsoCarla M CudaDavid A AllmanJames S RushCatherine H RegnierGrazyna WieczorekRita R AllowayAdele R ShieldsBrian M BakerE Steve WoodleDavid A HildemanPublished in: bioRxiv : the preprint server for biology (2023)
Bulk analysis of renal allograft biopsies (rBx) identified RNA transcripts associated with acute cellular rejection (ACR); however, these lacked cellular context critical to mechanistic understanding. We performed combined single cell RNA transcriptomic and TCRα/β sequencing on rBx from patients with ACR under differing immunosuppression (IS): tacrolimus, iscalimab, and belatacept. TCR analysis revealed a highly restricted CD8 + T cell clonal expansion (CD8 EXP ), independent of HLA mismatch or IS type. Subcloning of TCRα/β cDNAs from CD8 EXP into Jurkat76 cells (TCR -/- ) conferred alloreactivity by mixed lymphocyte reaction. scRNAseq analysis of CD8 EXP revealed effector, memory, and exhausted phenotypes that were influenced by IS type. Successful anti-rejection treatment decreased, but did not eliminate, CD8 EXP , while CD8 EXP were maintained during treatment-refractory rejection. Finally, most rBx-derived CD8 EXP were also observed in matching urine samples. Overall, our data define the clonal CD8 + T cell response to ACR, providing novel insights to improve detection, assessment, and treatment of rejection.
Keyphrases
- single cell
- regulatory t cells
- nk cells
- high throughput
- oxidative stress
- dendritic cells
- intensive care unit
- induced apoptosis
- electronic health record
- cell death
- deep learning
- big data
- respiratory failure
- peripheral blood
- endoplasmic reticulum stress
- quantum dots
- acute respiratory distress syndrome
- mechanical ventilation